Production of a human single-chain variable fragment antibody against esophageal carcinoma

AIM: To construct a phage display library of human singlechain variable fragment (scFv) antibodies associated with esophageal cancer and to preliminarily screen a scFvantibody against esophageal cancer.METHODS: Total RNA extracted from metastatic lymph nodes of esophageal cancer patients was used to construct a scFv gene library. Rescued by M13K07 helper phage, the scFv phage display library was constructed, esophageal cancer cell line Eca 109 and normal human esophageal epithelial cell line (NHEEC) were used for panning and subtractive panning of the scFv phage display library to obtain positive phage clones. Soluble scFv was expressed in E.coliHB2151 which was transfected with the positive phage clone, then purified by affinity chromatography.Relative molecular mass of soluble scFv was estimated by Western blotting, its bioactivity was detected by cell ELISA assay. Sequence of scFv was determined using the method of dideoxynucleotide sequencing.RESULTS: The size of scFv gene library was approximately 9×10^6 clones. After four rounds of panning with Eca109 and three rounds of subtractive panning with NHEEC cells,25 positive phage clones were obtained. Soluble scFv was found to have a molecular mass of 31 ku and was able to bind to Ecal09 cells, but not to HeLa and NHEEC cells.Variable heavy (VH) gene from one of the positive clones was shown to be derived from the γ chain subgroup IV of immunoglobulin, and variable light (VL) gene from the κ chain subgroup I of immunoglobulin.CONCLUSION: A human scFv phage display library can be constructed from the metastatic lymph nodes of esophageal cancer patients. A whole human scFv against esophageal cancer shows some bioactivity.

Association of HLA-B Alleles With Human Immunodeficiency Virus Type 1 Infection in the Yi Ethnic Group in Sichuan Province

Objective To determine the distribution of HLA-B alleles in the Chinese Yi ethnic group and its association with HIV infection. Methods One hundred and six unrelated healthy HIV negative and 73 HIV positive Chinese Yi ethnic individuals were typed by PCR-SSP. Results The frequency of alleles B*07, B*35, and B*46 were increased in HTV-1 -positive subjects, whereas the alleles B*55, B*44 and B*78 were absent in the HIV-infected persons studied. The B*46 allele was present in a significantly higher gene frequency among HIV-1-positive individuals (P=0.02, OR=3.32, 95% CI=1.13-9.78) compared with control subjects. Conclusion HLA-B*46 may be associated with its susceptibility to HIV-1 infections.

Development of a semi-quantitative food frequency questionnaire for middle-aged inhabitants in the Chaoshan area, China

AIM:This paper aims to develop a data-based semi-quantitative food frequency questionnaire (SQFFQ) covering both urban and rural areas in the Chaoshan region of Guangdong Province, China, for the investigation of relationships between food intake and lifestyle-related diseases among middle-aged Chinese. METHODS: We recruited 417 subjects from the general population and performed an assessment of the diet, using a 3-d weighed dietary record survey. We employed contribution analysis (CA) and multiple regression analysis (MRA) to select food items covering up to a 90% contribution and a 0.90 R2, respectively. The total number of food items consumed was 523 (443 in the urban and 417 in the rural population) and the intake of 29 nutrients was calculated according to the actual consumption by foods/ recipes. RESULTS: The CA selected 233,194, and 183 foods/recipes for the combined, the urban and the rural areas, respectively, and then 196,157, and 160 were chosen by the MRA. Finally, 125 foods/recipes were selected for the final questionnaire. The frequencies were classified into eight categories and standard portion sizes were also calculated. CONCLUSION: For adoption of the area-specific SQFFQ, validity and reproducibility tests are now planned tc determine how the combined SQFFQ performs in actual assessment of disease risk and benefit.